繖花龍吐珠對綠膿桿菌之抗菌活性研究

The antibacterial activity of Hedyotis corymbosa (L.) Lam.against Pseudomonas aeruginosa

李民凱(Min-Kai Lee)

指導教授 ： 梁文俐

臺北醫學大學/藥學院/生藥學研究所/碩士(2006年)

摘要

綠膿桿菌（Pseudomonas aeruginosa）為造成院內感染的重要病原菌，容易造成的感染包括心內膜炎、細菌性肺炎、尿道感染和皮膚感染，當醫院內的癌症或燒燙傷病人被綠膿桿菌感染後，致死率高達50 %。為尋找新的抗菌物質以對抗致死率極高的菌株，在這個實驗中我們以瓊脂紙錠擴散法與連續稀釋法檢測56種中草藥後，發現繖花龍吐珠（Hedyotis corymbosa (L.) Lam.）粗萃物具有良好的抑菌活性。在殺菌時間曲線試驗結果顯示，當粗萃物劑量為1024 ug/ml併用抗生素ofloxacin 2 ug/ml和ciprofloxacin 0.25 ug/ml對於Pseudomonas aeruginosa ATCC 27853，於24小時內即具有99.9％殺菌效果。繖花龍吐珠經過管柱層析法進行成分分離，並配合活性追蹤，分離出三個有效成分，再經結構鑑定與抑菌活性評估，發現palmitic acid抑菌活性最佳。當palmitic acid在128 ug/ml的劑量下合併ofloxacin和ciprofloxacin，可使ofloxacin對ATCC 27853的最小抑菌濃度由2降至0.5 ug/ml，也可使ciprofloxacin對ATCC 27853的最小抑菌濃度由0.25降至0.065 ug/ml。另一方面利用穿透式電子顯微鏡觀察palmitic acid對Pseudomonas aeruginosa ATCC 27853的抑菌機制，發現其可以使細菌體中心產生空泡，我們進而推測這是palmitic acid傷害細菌體的方式。

關鍵字

綠膿桿菌、繖花龍吐珠、palmitic acid

並列摘要

Pseudomonas aeruginosa is becoming major pathogen and causing infections within the hospital. Infections caused by P. aeruginosa include endocarditis, bacteremia pneumonia, urinary tract infections, and skin infections. P. aeruginosa infection is a serious problem in patients hospitalized with cancer, and burns, the case fatality is 50 percent. In order to find new and more effective antibiotic against P. aeruginosa, disc agar diffusion method and series dilution test were used for testing the fifty-six antibacterial Chinese herbs. Result of our study, we found that Hedyotis corymbosa (L.) Lam. crude extract had the best antibacterial activity among fifty-six Chinese herbs. In time-kill cruve test, the crude extract of Hedyotis corymbosa (L.) Lam. of 1024 ug/ml combined with clinical antibiotics ofloxacin 2 ug/ml or ciprofloxacin 0.25 ug/ml which approached 99 % bactericidal activity within twenty four hours. Further more, we used column chromatography followed its antibacterial activity isolated three compounds from the crude extract, and determined the structures based on the analysis of NMR spectra. We found that palmitic acid have the best antibacterial activity. When palmitic acid 128 ug/ml combined with ofloxacin against P. aeruginosa ATCC 27853, the MIC of ofloxacin was falling from 2 to o.5 ug/ml. When palmitic acid 128 ug/ml combined with ciprofloxacin against P. aeruginosa ATCC 27853, the MIC of ciprofloxacin was falling from 0.25 to 0.0625 ug/ml. Otherwise we used TEM to observe its mechanism of action. We found that P. aeruginosa ATCC 27853 treated palmitic acid forming bleb. We speculated that is the way palmitic acid damaged P. aeruginosa ATCC 27853.

並列關鍵字

Pseudomonas aeruginosa, Hedyotis corymbosa (L.) Lam., palmitic acid

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